噬菌体LH-01治疗不同宿主菌所致小鼠致死性败血症影响因素研究

发布时间：2018-06-29 21:27

本文选题：肺炎克雷伯菌 + 噬菌体治疗　；参考：《中国病原生物学杂志》2015年05期

【摘要】：目的观察肺炎克雷伯菌噬菌体LH-01治疗不同宿主菌所致小鼠败血症的效果,分析影响疗效因素,探索评价噬菌体疗效的方法。方法通过比较噬菌斑和成斑率,从9株LH-01宿主菌中选择对其敏感性较强的菌株。尾静脉注射绝对致死量的不同敏感菌致小鼠败血症,2h后通过尾静脉注射LH-01治疗并观察其疗效。比较LH-01在小鼠新鲜血清、补体热灭活血清,以及在标准补体溶血活性由低至高的溶液中对敏感菌的杀灭能力,明确引起LH-01对不同敏感菌感染组小鼠疗效差异的因素。结果 9株宿主菌中KP6、KP13、KP16和KP22在营养肉汤中对LH-01最为敏感。当小鼠全身感染致死量的这4种细菌后,LH-01的治疗可使KP6感染组小鼠生存率达90%(9/10),KP13组为30%(3/10),而KP16组和KP22组小鼠全部死亡。体外试验显示,除LH-01对KP6的作用不受补体影响外,LH-01对其余3株菌的杀灭作用均受补体影响,其中25U/ml的补体活性即可完全抑制LH-01对KP16和KP22的杀灭作用,而完全抑制对KP13的杀灭作用所需的补体活性≥50U/ml。结论补体是影响LH-01治疗小鼠肺炎克雷伯菌感染疗效的最主要原因。血清中噬菌体对致病菌的杀菌力和补体干扰杀菌的程度有助于预测噬菌体疗效,指导细菌感染的噬菌体治疗。
[Abstract]:Objective to observe the efficacy of Klebsiella pneumoniae bacteriophage LH-01 in the treatment of septicemia in mice induced by different host bacteria, analyze the influencing factors and explore the method of evaluating the efficacy of bacteriophage. Methods by comparing plaque and plaque forming rate, 9 strains of LH-01 were selected. Mice with septicemia were treated by tail vein injection of LH-01 2 hours after injection of different sensitive bacteria with absolute lethal dose and the curative effect was observed. To compare the killing ability of LH-01 against sensitive bacteria in fresh serum, heat-inactivated serum and standard complement hemolytic activity from low to high solution, the factors that caused the difference of the efficacy of LH-01 to mice infected with different sensitive bacteria were determined. Results KP6, KP13, KP16 and KP22 were most sensitive to LH-01 in nutrient broth. The survival rate of mice in KP6 infected group was 90% (9 / 10), and that in KP13 group was 30% (3 / 10), while that in KP16 and KP22 groups were all dead. In vitro experiments showed that the killing effect of LH-01 on KP6 was not affected by complement, and the cytotoxicity of LH-01 to KP16 and KP22 could be completely inhibited by the complement activity of 25 U / ml, and the effect of LH-01 on KP16 and KP22 was completely inhibited by the addition of LH-01 to KP16 and KP22, and the effect of LH-01 on KP16 and KP22 was inhibited by LH-01. The complement activity required to completely inhibit the killing effect of KP13 was more than 50 U / ml. Conclusion complement is the most important factor affecting the efficacy of LH-01 in the treatment of Klebsiella pneumoniae infection in mice. The bactericidal ability of bacteriophages in serum and the degree of complement interference in bactericidal efficacy can be used to predict the efficacy of bacteriophage and to guide the bacteriophage therapy of bacterial infection.
【作者单位】：河南医学高等专科学校病原生物学与免疫学教研室;郑州大学基础医学院病原生物学教研室;
【基金】：河南省科技厅重点科技攻关项目(No.142102310440) 河南省医学科技攻关计划项目(No.201303199)
【分类号】：R515.3

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